| 摘要: |
| 以中间球海胆(Strongylocentrotus intermedius)作为实验对象,研究其在高温胁迫条件下热休克蛋白基因中hsp70基因和hsp90基因(以下简称hsp70和hsp90)的响应。采用荧光实时定量PCR技术,检测hsp70和hsp90在40枚中间球海胆的不同组织(管足、体腔细胞、肠、性腺和口器)中的相对表达量,以及在海胆耐热家系和对照家系(共240枚)的管足组织中的相对表达量。结果显示:(1)中间球海胆的各组织中均有hsp70和hsp90基因的表达,且对温度急剧变化均有应答。高温诱导过程中,海胆管足、体腔细胞和肠组织中hsp70和hsp90基因的相对表达量经过6h(29℃)达到最大值且显著高于其初始值(P<0.05);性腺中hsp70和hsp90基因的相对表达量分别经过12h(24℃)和24h(22℃)达到最大值;口器中两个基因的相对表达量都经过12h达到最大值。(2)高温诱导过程中, hsp70和hsp90基因在海胆体腔细胞和管足中的表达趋势相似,且表达量在各时间点的相对差异不显著(P>0.05)。(3)高温诱导过程中, hsp70基因在海胆耐热家系和对照家系中的表达趋势均为先升高后降低,且耐热家系和对照家系间各时间点的相对表达量没有显著差异(P>0.05); hsp90基因的表达量随温度的变化呈现先升高后降低的趋势,诱导12h时在2个耐热家系中的相对表达量显著高于2个对照家系(P<0.05)。结果表明,温度变化会显著影响中间球海胆hsp90基因的表达, hsp90基因可作为中间球海胆耐热品系选育的候选指示因子。 |
| 关键词: 中间球海胆 hsp70 hsp90 温度 实时荧光定量PCR |
| DOI：10.11693/hyhz20141200366 |
| 分类号: |
| 基金项目:国家高技术研究发展计划(863)重大项目,2012AA10A412号;辽宁省农业攻关及成果产业化项目,2015203003号。 |
附件 |
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| RELATIVE EXPRESSION OF GENES HSP70 AND HSP90 IN SEA URCHIN STRONGYLOCENTROTUS INTERMEDIUS IN THERMAL STRESS |
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BAI Xue-Qiu, PANG Zhen-Guo, ZHANG Wei-Jie, CHANG Ya-Qing, GAO Yin-Xue, DING Jun
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Key Laboratory of Mariculture & Stock Enhancement in North China's Sea, Ministry of Agriculture, Dalian Ocean University, Dalian 116023, China
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| Abstract: |
| To understand the response behavior of heat shock proteins(hsp70 and hsp90) under thermal stress in sea urchin Strongylocentrotus intermedius, we determined the relative expression quantity of hsp70 and hsp90 in adult urchin tissues(including tube-feet, coelomocytes, intestine, gonad and lantern using the RT-PCR(reverse transcription polymerase chain reaction). Different expressions of the two genes between thermal-tolerant chains and control ones were compared especially in tube feet. The results show that the two genes are expressed ubiquitously in various adult tissues and responded quickly to the thermal stress. Relative expressions of the genes in tube feet, coelomocytes, and intestine of sea urchins reached their highest levels in 6h(29℃), which is significantly higher than the control points(P<0.05). The relative expression of hsp70 and hsp90 reached the maximum in 12h(24℃) and 24h(22℃) respectively in gonads. Relative expression of the two genes reached maximum at 12h in lantern. In tube feet and coelomocytes, the expression trends of hsp70 and hsp90 were quite similar and no significant difference was found in different time(P>0.05). The expression of hsp70 increased gradually from the beginning of experiment, then decreased despite of temperature variation, and no significant difference between heat-resistant and control sea urchin chains at all the time points(P>0.05). In heat-resistant chains, the expression of hsp90 increased at first, and then decreased gradually at 12h, but the expression of hsp90 in the two heat-resistant chains were higher than the two control ones(P<0.05). The result indicates that thermal stress can significantly affect the expression of hsp90; and therefore, hsp90 can be used as an internal thermal-tolerant indicator in heat-resistant selection breeding of S. intermedius. |
| Key words: Strongylocentrotus intermedius hsp70 hsp90 temperature RT-PCR |
